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Using Affymetrix porcine Gene-Chip analyses, we found that Dickkopf2 ( DKK2 ), a WNT antagonist, is differentially expressed in pre-ovulatory follicles between Large White and Chinese Taihu sows. This study aims to identify the regulatory factors responsible for  DKK2  expression. Deletion fragment and mutation analyses identified  DKK2-D3  as the porcine  DKK2  core promoter. There were four C/EBPβ binding sites within the  DKK2  core promoter. The C allele that results from a spontaneous alteration ( DKK2  c.−1130 T > C) in the core promoter was associated with a higher total number born (TNB) and a higher number born alive (NBA) in all parities in a synthetic pig population. This was possibly the result of a change in C/EBPβ binding ability, which was confirmed using chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays (EMSA). Moreover, C/EBPβ specifically bound to and activated the  DKK2  promoter, as revealed by mutation analysis, overexpression and RNA interference (RNAi) experiments. We also confirmed that miR-27a is a negative regulator of the  DKK2  gene using miR-27a overexpression and inhibition experiments and mutation analyses. RTCA xCELLigence experiments showed that miR-27a suppressed Chinese hamster ovary (CHO) cell proliferation by down-regulating  DKK2  gene expression. Taken together, our findings suggest that C/EBPβ and miR-27a control  DKK2  transcription.

The transcription factor ccaat/enhancer binding protein β (C/EBPβ) and miR-27a regulate the expression of porcine Dickkopf2 (DKK2)